• Archives of Pharmacal Research
• /
• 제22권3호
• /
• pp.274-278
• /
• 1999

Expression of the active urease of the enterobacterium, Klebsiella aerogens, requires the presence of the accessory genes (ureD, ureE, ureF, and ureG) in addition to the three structural genes (ureA, ureB, and ureC). These accessory genes are involved in functional assembly of the nickel-metallocenter for the enzyme. Characterization of ureF gene has been hindered, however, since the UreF protein is produced in only minute amount compared to other urease gene products. In order to overexpress the ureF gene, a recombinant pMAL-UreF plasmid was constructed from which the UreF was produced as a fusion with maltose-binding protein. The MBP-UreF fusion protein was purified by using an amylose-affinity column chromatography followed by an anion exchange column chromatography. Polyclonal antibodies raised against the fusion protein were purified and shown to specifically recognize both MBP and UreF peptides. The UreF protein was shown to be unstable when separated from MBP by digestion with factor Xa.

• Asian-Australasian Journal of Animal Sciences
• /
• 제20권9호
• /
• pp.1342-1348
• /
• 2007

POU1F1 is a positive regulator for GH, PRL and TSH${\beta}$ and its mutations associate with production traits in ruminant animals. We described a DdeI PCR-RFLP method for detecting a silent allele in the goat POU1F1 gene: TCT (241Ser)>TCG (241Ser). Frequencies of $D_1$ allele varied from 0.600 to 1.000 in Chinese 801 goats. Significant associations of DdeI polymorphism with production traits were found in milk yield (*p<0.05), litter size (*p<0.05) and one-year-old weight (*p<0.05) between different genotypes. Individuals with genotype $D_1D_1$ had a superior performances when compared to those with genotype $D_1D_2$ (*p<0.05). Hence, the POU1F1 gene was suggested to the potential candidate gene for superior milk performance, reproduction trait and weight trait. Genotype $D_1D_1$, characterized by a DdeI PCR-RFLP detection, was recommended to geneticists and breeders as a molecular marker for better performance in the goat industry.

• Fisheries and Aquatic Sciences
• /
• 제15권4호
• /
• pp.325-335
• /
• 2012

The present study characterized the fast skeletal myosin light chain-2 gene (mlc2f) in the euryhaline Javanese ricefish (Oryzias javanicus: Beloniformes). Coding nucleotide and deduced amino acid sequences of Javanese ricefish mlc2f were well conserved in the vertebrate lineage. Javanese ricefish mlc2f showed a typical seven-exon structure, and its promoter exhibited transcription factor binding motifs common to most muscle-specific genes. However, Javanese ricefish mlc2f also displayed tandem duplications of intronic sequences in both intron 1 and intron 3. Based on quantitative reverse transcription-polymerase chain reaction, the mlc2f transcripts were highly predominant in skeletal muscles of adults and were differentially modulated during embryonic development. Microinjection of the mlc2f promoter-driven red fluorescent protein (RFP) reporter construct successfully exhibited heterologous expression of the fluorescent reporter, primarily in muscular areas of hatchlings, although the distribution pattern of RFP signals was not uniform due to the mosaic nature of the introduced transgene. Data from this study indicate that the Javanese ricefish mlc2f gene has undergone "intra-intronic" duplication events in a species-specific manner and that the mlc2f regulator may also be useful in heterologous expression assays of the skeletal muscles of this species.

• Journal of Microbiology and Biotechnology
• /
• 제32권12호
• /
• pp.1527-1536
• /
• 2022

Escherichia coli can use allantoin as its sole nitrogen source under anaerobic conditions. The ureidoglycolate produced by double release of ammonia from allantoin can flow into either the glyoxylate shunt or further catabolic transcarbamoylation. Although the former pathway is well studied, the genes of the latter (catabolic) pathway are not known. In the catabolic pathway, ureidoglycolate is finally converted to carbamoyl phosphate (CP) and oxamate, and then CP is dephosphorylated to carbamate by a catabolic carbamate kinase (CK), whereby ATP is formed. We identified the ybcF gene in a gene cluster containing fdrA-ylbE-ylbF-ybcF that is located downstream of the allDCE-operon. Reverse transcription PCR of total mRNA confirmed that the genes fdrA, ylbE, ylbF, and ybcF are co-transcribed. Deletion of ybcF caused only a slight increase in metabolic flow into the glyoxylate pathway, probably because CP was used to de novo synthesize pyrimidine and arginine. The activity of the catabolic CK was analyzed using purified YbcF protein. The V_max is 1.82 U/mg YbcF for CP and 1.94 U/mg YbcF for ADP, and the K_M value is 0.47 mM for CP and 0.43 mM for ADP. With these results, it was experimentally revealed that the ybcF gene of E. coli encodes catabolic CK, which completes anaerobic allantoin degradation through substrate-level phosphorylation. Therefore, we suggest renaming the ybcF gene as allK.

• Animal Systematics, Evolution and Diversity
• /
• 제22권1호
• /
• pp.87-89
• /
• 2006

The mitochondrial cytochrome b gene of the Korean subspecies (Falco tinnunculus interstinctus) of the common kestrel has been analyzed. According to the molecular phylogeny of six subspecies of common kestrel, six subspecies of the common kestrel were divided into two clades: the first clade is composed of the South African subspecies (F. t. rupicolus) and the second clade includes 5 subspecies (F. t. tinnunculus, F. t. rufescens, F. t. interstinctus, F. t. canariensis and F. t. dacotiae) of the common kestrel. Korean subspecies, F. t. interstinctus was closely related to F. t. rufescens and original subspecies F. t. tinnunculus.

• 한국가축번식학회지
• /
• 제25권1호
• /
• pp.85-90
• /
• 2001

본 연구는 growth hormone receptor(GHR) gene의 동물생리에 미치는 영향을 연구하기 위해 metallothionein promoter와 GHR gene을 이용하여 생쥐의 1-cell 수정란에 DNA 미세주입법에 의해 형질전환생쥐를 생산하였다. 세마리의 형질전환생쥐가 생산되었는데 DNA 분석결과 4~8 copy의 GHR 유전자를 지닌 것으로 확인되었다 이들 세 마리의 GHR 형질전환생쥐를 정상 형질전환생쥐와 교미시켜 F$_1$과 F$_2$ 새끼를 생산하였는데 이들의 전달율은 F$_1$에서 20~50%였고 F$_2$에서는 약 50%를 나타내어 모자익형태로 유전자가 정착되었음을 확인할 수 있었다. 3주령까지의 사망률은 Fl과 F2 새끼에서 약 10~30%를 나타내어 GHR유전자의 발현이형질전환생쥐의 초기 사망에 영향을 미치는 것으로 나타났다.

• 방사선산업학회지
• /
• 제2권3호
• /
• pp.121-128
• /
• 2008

Flavanone-3-hydroxylase (F3H) is one of the key enzymes for the biosynthesis of flavonals, anthocyanins, catechins and proanthocyanins. F3H catalyzes the $3{\beta}$-hydroxylation of (2S)-flavonones to form (2R, 3R)-dihydroflavonols. In this report, we isolated a full-length cDNA of RocF3H from black raspberry (Rubus occidentalis L.) using a reverse transcriptase-PCR and rapid amplification of the cDNA ends (RACE)-PCR. The full-length cDNA of RocF3H contains a 1,098 bp open reading frame (ORF) encoding a 365 amino acid protein with a calculated molecular weight of about 41.1 kDa and isoelectric point (pI) of 5.45. The genomic DNA analysis revealed that the RocF3H gene had three exons and two introns. Comparison of the deduced amino acid sequence of the RocF3H with other F3Hs revealed that the protein is highly homologous with various plant species. The conserved amino acids ligating the ferrous iron and the residues participating in the 2-oxoglutarate binding (R-X-S) were found in RocF3H at the similar positions to other F3Hs. Southern blot analysis indicated that RocF3H exist a multi-gene family. The isolation of RocF3H gene will be helpful to further study the role of F3H gene in the biosynthesis of flavonoids in R. occidnetalis.

• Asian-Australasian Journal of Animal Sciences
• /
• 제21권4호
• /
• pp.489-493
• /
• 2008

In 2004, Jorgensen and coworkers proposed the MUC4 gene as a candidate gene of enterotoxigenic Escherichia coli (ETEC) F4ab/ac receptor in piglets and a mutation of $G{\rightarrow}C$ in intron 7 of MUC4 was identified to be associated with the ETEC F4ab/ac adhesion phenotypes. In this study, we used 310 piglets of three breeds (Landrace, Large White and Chinese Songliao Black) to analyze the relationship between this mutation and the F4ab/ac adhesion phenotype. The results show that the genotypes at this site and the ETEC F4ab/ac adhesion phenotypes were not completely consistent, although they are very strongly associated. Among the individuals with genotype CC, which was identified as a resistant genotype to F4ab/ac adhesion, only 72.1% (124/172) were non-adhesive to ETEC F4ab and 77.9% (134/172) were non-adhesive to ETEC F4ac infections. This suggests that this mutation may not be the causative mutation for ETEC F4ab/ac adhesion, rather, the actual causative mutation may be in another gene closely linked to MUC4, or at aother site within the MUC4 gene. Our results also suggest that the receptors of F4ab and F4ac may be determined by two different but closely linked loci. In order to screen other genes related to F4ab/ac adhesion in piglets, the mRNA profiles from six full sib piglets, of which three were adhesive to ETEC F4ab/ac and three non-adhesive, were analyzed by suppression subtractive hybridization (SSH). One up-regulated gene, Ep-CAM, was selected for further analysis based on its role in the intestinal epithelial cells adhesion. Using real-time RT-PCR, we found that the Ep-CAM gene was significantly up-regulated in the piglets adhesive to F4ab/ac. It was mapped to SSC3q11-q14 by radiation hybrid mapping.

• 대한한방내과학회지
• /
• 제31권1호
• /
• pp.25-39
• /
• 2010

Background : At high glucose levels in $\beta$-cells, cell viability and insulin secretion are decreased by glucotoxicity. Sopyung-tang(SPT) had an effect on blood glucose level decrease and antioxidant enzyme activities in streptozotocin-induced diabetic rats. Objectives : This study performed a series of experiment to verify the effects of SPT extract on the cell viability, antioxidant enzyme activities, insulin secretion and insulin mRNA expression at hyperglycemic states of RIN-m5F. Methods : After treatment at various concentrations of SPT added to the RIN-m5F cells, cell viability by MTT assay, free radical-scavenging activity, SOD activity and insulin secretion were measured. Additionally, insulin-related gene expression was measured using real-time RT-PCR. Results : Compared to the control group, SPT extract showed considerable effects on RIN-m5F cell viability, DPPH radical-scavenging activity, superoxide dismutase (SOD) activity, insulin secretion and insulin-related gene expression. Conclusions : This study showed that SPT extract has an effect on $\beta$-cell cell viability, insulin secretion and insulin-related gene expression. Thus, SPT extract may be used for treatment of diabetes and its complications. Further mechanism studies of SPT seem to be necessary on the glucotoxicity and oxidative stress.

• Journal of Oral Medicine and Pain
• /
• 제24권2호
• /
• pp.219-234
• /
• 1999

치석에는 박리상피세포, 백혈구 등이 포함되어 있어 이들의 핵 내에 있는 DNA의 유전자형을 찾아내 개인식별을 할 수 있을 것으로 추정된다. 본 연구에서는 치석만으로 개인식별이 가능한지를 알아보고자 40명으로부터 채취한 치석을 증류수에 세척한 군과 세척하지 않은 군으로 나누어 DNA를 추출하고 중합효소연쇄반응법을 이용하여 증폭절편다형(AMP-FLPs)을 실시한 후 성별검사를 위한 X-Y homologous amelogenin gene과 유전자지문검사를 위한 STR유전좌위 LPL, F13B, Triplex(F13A01, FESFPS, vWA) 등 6개의 유전자를 검색하여 - X-Y homologous amelogenin gene과 LPL, F13B는 각각 증폭하였으며 F13A01, FESFPS, vWA 세 유전자는 동시에 증폭하였음 - 다음과 같은 결과를 얻었다. 1) X-Y homologous amelogenin gene 검색으로 세척군에서 27개의 검체 중 8개, 비세척군에서 13개 중 11개에서 성별검사가 가능하였다. 2) LPL유전자는 세척군, 비세척군에서 각각 27개 검체중 2개, 13개 검체 중 5개가 검색되었으며 3개의 대립유전자(10, 11, 12)와 4개의 유전자형 (10-10, 10-11, 10-12, 11-12)이 나타났다. 3) F13B유전자는 세척군, 비세척군에서 각각 27개 검체 중 1개, 13개 검체 중 4개가 검색되었으며 2개의 대립유전자(9, 10)와 2개의 유전자형(9-10, 10-10)을 관찰하였다. 4) F13A01유전자는 비세척군에서만 13개 검체 중 3개가 검색되었고 3개의 대립유전자(3.2, 4, 6)와 3개의 유전자형(3.2-3.2, 4-5, 4-6)을 관찰하였고, 세척군에서는 나타나지 않았다. 5) FESFPS유전자는 비세척군에서만 13개 검체 중 1개가 검색되었고 유전자 형은 11-12로 나타났다. 6) vWA유전자는 세척군, 비세척군에서 각각 1개씩 검색되었으며, 3개의 대립유전자형(14, 16, 17)와 2개의 유전자형(14-16, 14-17)을 관찰하였다. 이상의 결과를 종합해 볼 때, 치석은 X-Y homologous amelogenin gene증폭을 통한 성별검사와 단일 STR유전좌위 증폭을 통한 유전자지문형 검사에는 유용하나 복합 STR유전좌위의 검색에는 부적합한 것으로 나타났으며 법의학적시료로 응용이 가능한 것으로 사료된다.